Crystallization Control of Thermal Stability and Morphology of Hen Egg White Lysozyme Crystals by Ionic Liquids

Abstract

Ionic liquids (ILs) exhibit a variety of properties that make them attractive additives for biomaterials. 1-Butyl-3-methylimidazolium tetrafluoroborate ([C(4)mim]BF(4)), 1-butyl-3-methylimidazolium chloride ([C(4)mim]Cl), 1-butyl-3-methylimidazolium bromide ([C(4)mim]Br), and 1,3-dimethylimidazolium iodine([bmim]I), as additives during lysozyme crystallization, were tested for their effects on the thermal stability and morphology of lysozyme crystals obtained. [C(4)mim]Cl was chosen to evaluate the effect of IL addition concentration on the thermal stability of lysozyme. It is shown that the characteristic peak temperature and endothermic enthalpy values (DeltaH) for denaturation increase with increasing addition concentration. As for the degradation, peak temperatures decrease, whereas endothermic enthalpy values markedly increase with the rise of [C(4)mim]Cl addition concentration. In the case of adding [C(4)mim]BF(4), [C(4)mim]Br, and [bmim]I, similar thermal behaviors of lysozyme crystals were observed. The effect of ILs on thermal behaviors of lysozyme can be attributed to enhancing crystal contacts, changing conformational stability, or interaction among molecules, as evidenced by difference in crystal growth morphology. This study is especially helpful in controlling the thermal stability of lysozyme crystals and in gaining initial insight into potential crystallization conditions for prescreening ILs that stabilize the protein and other macromolecule crystals.